Eukaryotic DNA transposons can be classified into distinct superfamilies (Kapitonov & Jurka, 2004). One of the most widely distributed is the so-called hAT superfamily, which has active members in plants and insects. We began our structural studies of eukaryotic DNA transposases with Hermes, a hAT transposon that is active not only in the house fly from which it was isolated but also in other insects such as Aedes aegypti (Sarkar et al., 1997), the mosquito species that transmits yellow fever. A close relative of Hermes, the Herves transposon, is active in the malaria vector Anopheles gambiae (Arensburger et al., 2005). An active insect transposon is particularly interesting because it offers the potential to produce transgenic insects for controlling medically significant pests. Hermes transposition has been recapitulated in vitro and shown to employ a mechanism in which excision is accompanied by hairpin formation on the DNA flanking the transposon (Zhou et al., 2004), as also seen for the RAG1/2 recombinase of the adaptive immune system. In 2005, we determined the structure of an N-terminally truncated version of the 612-residue Hermes protein (Hickman et al., 2005). Hermes was shown to be a multidomain protein organized around the RNase H-like catalytic core characteristic of DDE transposases. The DDE catalytic core is disrupted by a large insertion domain whose presence conforms to the trend that DDE transposases capable of forming hairpins on their DNA substrates require an ancillary domain to provide the amino acids needed to promote hairpin formation and to stabilize them. In these initial studies, we crystallized only a proteolyzed dimeric form of Hermes, and we used our structural results to propose a model for the larger active assembly which appears to be hexameric. Our current focus is the full-length protein and its complexes with DNA. We have recently determined the structure of a catalytically active version of Hermes bound to its transposon ends to 3.3 A resolution. We are currently pursuing in vivo and in vitro studies to understand the implications of the complex we observe. More recently, we have also solved the structure of a hyperactive dimer of Hermes in complex with transposon ends and the adjacent flanking sequence. Other DNA transposition systems of interest to us include those that function in mammalian cells such as Tol2 from the medaka fish and piggyBac, an active moth transposon (Wu et al., 2006; Mitra et al., 2008). Dupuy, A.J., Akagi, K., Largaespada, D.A., Copeland, N.G., and Jenkins, N.A. (2005) Nature 436, 221-226. Hickman, A.B., et al. (2005) Nat. Struct. Mol. Biol. 12, 715-721. Wu, S.C., et al. (2006) Proc. Natl. Acad. Sci. USA 103, 15008-15013. Kapitonov, V.V. and Jurka, J. (2004) DNA Cell Biol. 23, 311-324. Mitra, R., Fain-Thornton, J., and Craig, N.L. (2008) EMBO J. 27, 1097-1109. Sarkar, A., Yardley, K., Atkinson, P.W., James, A.A., and O'Brochta, D.A. (1997) Insect Biochem. Mol. Biol. 27, 359-363. Zhou L.Q., et al. (2004) Nature 432, 995-1001.